Compounds and compositions for delivering active agents

ABSTRACT

Carrier compounds and compositions therewith which are useful in the delivery of active agents are provided Methods of administration and preparation are provided as well.

This application is a national phase application under 35 U.S.C. §371 ofInternational Application No. PCT/US99/17090, filed Jul. 27, 1999, whichwas published in English on Feb. 10, 2000 as International PublicationNo. WO 00/06534 and claims the benefit of U.S. Provisional ApplicationNo. 60/094,267, filed Jul. 27, 1998, and U.S. Provisional ApplicationNo. 60/104,466, filed Oct. 16, 1998.

FIELD OF THE INVENTION

The present invention relates to compounds for delivering active agents,and particularly biologically or chemically active agents. Thesecompounds are used as carriers to facilitate the delivery of an activeagent to a target. The carrier compounds are well suited to formnon-covalent mixtures with biologically-active agents for pulmonary,oral, subcutaneous, intranasal, sublingual, buccal, ocular,intracolonic, intraduodenal, rectal, vaginal, mucosal, transdermal,intradermal, parenteral, intravenous, and intramuscular administrationto animals, as well as crossing the blood-brain barrier. Methods for thepreparation and administration of such compositions are also disclosed.

BACKGROUND OF THE INVENTION

Conventional means for delivering active agents are often severelylimited by biological, chemical, and physical barriers. Typically, thesebarriers are imposed by the environment through which delivery occurs,the environment of the target for delivery, or the target itself.Biologically or chemically active agents are particularly vulnerable tosuch barriers.

For example, in the delivery to animals of biologically active orchemically active pharmacological and therapeutic agents, barriers areimposed by the body. Examples of physical barriers are the skin andvarious organ membranes that must be traversed before reaching a target.Chemical barriers include, but are not limited to, pH variations, lipidbi-layers, and degrading enzymes.

These barriers are of particular significance in the design of drugdelivery systems. For example, oral delivery of many biologically orchemically active agents would be alternate routes of choice foradministration to animals if not for biological, chemical, and physicalbarriers such as varying pH in the gastro-intestinal membranes. Amongthe numerous agents which are not typically amenable to oraladministration are biologically or chemically active peptides, such ascalcitonin and insulin; polysaccharides, and in particularmucopolysaccharides including, but not limited to, heparin; heparinoids;antibiotics; and other organic substances. These agents are rapidlyrendered ineffective or are destroyed in the gastro-intestinal tract byacid hydrolysis, enzymes, and the like. In addition, the size andstructure of macromolecular drugs may prohibit absorption.

Crossing the blood-brain barrier, as well as oral, subcutaneous,intranasal, sublingual, buccal, ocular, intracolonic, intraduodenal,mucosal, transdermnal or pulmonary delivery to the circulatory systemfor many biologically active agents could be the route of choice foradministration to animals if not for physical barriers such as the skin,lipid bi-layers, and various organ membranes that are relativelyimpermeable to certain biologically active agents, but one or more ofwhich must be traversed before an agent delivered via these routes canreach the circulatory system. Additionally, delivery such as, forexample, sublingual delivery may be impeded by chemical barriers such asthe varying pH in the gastrointestinal (GI) tract and the presence ofpowerful digestive enzymes.

Earlier methods for orally administering vulnerable pharmacologicalagents have relied on the co-administration of adjuvants (e.g.,resorcinols and non-ionic surfactants such as polyoxyethylene oleylether and n-hexadecylpolyethylene ether) to increase artificially thepermeability of the intestinal walls, as well as the co-administrationof enzymatic inhibitors (e.g., pancreatic trypsin inhibitors,diisopropylfluorophosphate (DFF) and trasylol) to inhibit enzymaticdegradation.

Liposomes have also been described as drug delivery systems for insulinand heparin. See, for example, U.S. Pat. No. 4,239,754; Patel et al.(1976), FEBS Letters, Vol. 62, pg. 60; and Hashimoto et al. (1979),Endocrinology Japan, Vol. 26, pg. 337.

However, broad spectrum use of such drug delivery systems is precludedbecause: (1) the systems require toxic amounts of adjuvants orinhibitors; (2) suitable low molecular weight cargos, i.e. activeagents, are not available; (3) the systems exhibit poor stability andinadequate shelf life; (4) the systems are difficult to manufacture; (5)the systems fail to protect the active agent (cargo); (6) the systemsadversely alter the active agent; or (7) the systems fail to allow orpromote absorption of the active agent.

More recently, microspheres of artificial polymers of mixed amino acids(proteinoids) have been used to deliver pharmaceuticals. For example,U.S. Pat. No. 4,925,673 describes drug-containing proteinoid microspherecarriers as well as methods for their preparation and use. Theseproteinoid microspheres are useful for the delivery of a number ofactive agents.

However, there is still a need for simple, inexpensive delivery systemswhich are easily prepared and which can deliver a broad range of activeagents by various routes.

SUMMARY OF THE INVENTION

The present invention provides compounds having the formula

and salts thereof. These compounds are useful in the delivery of activeagents.

Another embodiment of the invention is a composition comprising:

(A) at least one active agent; and

(B) a carrier comprising a compound having the formula

 or a salt thereof.

Also provided are dosage unit forms comprising the composition above andan excipient, a diluent, a disintegrant, a lubricant, a plasticizer, acolorant, a dosing vehicle, or any combination thereof.

Methods for administering a biologically-active agent to an animal inneed of the agent by the pulmonary, oral, subcutaneous, intranasal,sublingual, buccal, ocular, intracolonic, intraduodenal, rectal,vaginal, mucosal, transdermal, intradermal, parenteral, intravenous, orintramuscular route or crossing the blood-brain barrier with thecompositions of the present invention are also provided.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graphical illustration of blood glucose levels afterpulmonary delivery of insulin.

FIG. 2 is a graphical illustration of blood glucose levels afterpulmonary delivery of insulin.

DETAILED DESCRIPTION OF THE INVENTION

One group of compositions of the present invention include an activeagent and a carrier which includes the compound2-(4-(N-salicyloyl)aminophenyl)propionic acid or salts thereof, such asthe sodium salt thereof (sodium2-(4-(N-salicyloyl)aminophenyl)propionate). These compositions may beused to deliver various active agents through various biological,chemical, and physical barriers and are particularly suited fordelivering active agents which are subject to environmental degradation.The compositions of the subject invention are particularly useful fordelivering or administering biologically or chemically active agents toany animals such as birds, including, but not limited to, chickens;mammals, such as primates, and particularly humans; and insects.

Other advantages of the present invention include the use of easy toprepare, inexpensive raw materials. The compositions and the formulationmethods of the present invention are cost effective, simple to perform,and amenable to industrial scale up for commercial production.

Pulmonary, oral, subcutaneous, intranasal, sublingual, buccal, ocular,intracolonic, intraduodenal, rectal, vaginal, mucosal, transdermal,intradermal, parenteral, intravenous, and intramuscular coadministrationof an active agent, such as, for example, recombinant human growthhormone (rhGH); salmon calcitonin; insulin heparin, including, but notlimited to, low molecular weight heparin and very low molecular weightheparin; parathyroid hormone (PTH); and compounds in compositions asdescribed herein result in an increased bioavailability of the activeagent compared to administration of the active agent alone.

Active Agents

Active agents suitable for use in the present invention includebiologically and chemically active agents including, but not limited to,cosmetics, pesticides, pharmacological agents, and therapeutic agents.For example, biologically or chemically active agents suitable for usein the present invention include, but are not limited to, proteins;polypeptides; peptides, and particularly small peptides; hormones, andparticularly hormones which by themselves do not or only a fraction ofthe administered dose passes through the gastro-intestinal mucosa and/orare susceptible to chemical cleavage by acids and enzymes in thegastro-intestinal tract; polysaccharides, and particularly mixtures ofmuch-polysaccharides; carbohydrates; lipids; other organic compounds; orany combination thereof. Further examples include, but are not limitedto, the following, including synthetic, natural or recombinant sourcesthereof: growth hormones, including human growth hormones (hGH),recombinant human growth hormones (rhGH), bovine growth hormones, andporcine growth hormones; growth hormone-releasing hormones; interferons,including α, β and γ; interleukin-1; interleukin-II;insulin;insulin-like growth factor, including IGF-1; heparin, includingunfractionated heparin, heparinoids, dermatans, chondroitins, lowmolecular weight heparin, very low molecular weight heparin and ultralow molecular weight heparin; calcitonin, including salmon, eel andhuman; erythropoietin (EPO); atrial naturetic factor; antigens;monoclonal antibodies; somatostatin; protease inhibitors;adrenocorticotropin, gonadotropin releasing hormone; oxytocin;leutinizing-hormone-releasing-hormone; follicle stimulating hormone;glucocerebrosidase; thrombopoietin; filgrastim; prostaglandins;cyclosporin; vasopressin; cromolyn sodium (sodium or disodiumchromoglycate); vancomycin; desferrioxamine (DFO); parathyroid hormone(PTH), including its fragments; antimicrobials, including anti-fungalagents; analogs, fragments, mimetics or polyethylene glycol(PEG)-modified derivatives of these compounds; or any combinationthereof.

Carriers

The carrier compounds of the present invention include

2-(4-(N-salicyloyl)aminophenyl)propionic acid

and salts thereof. Salts of the compound include, but are not limitedto, organic or inorganic salts, such as the sodium salt:

Sodium-2-(4-(N-salicyloyl)aminophenyl)propionate

In addition, carriers of the present invention include poly amino acidsand peptides comprising one or more of the carrier compounds of thepresent invention.

These carrier compounds, poly amino acids, and peptides may be used todeliver active agents including, but not limited to, biologically orchemically active agents, such as for example, pharmacological andtherapeutic agents.

An amino acid is any carboxylic acid having at least one free aminegroup and includes naturally occurring and synthetic amino acids.

Poly amino acids are either peptides or two or more amino acids linkedby a bond formed by other groups which can be linked, e.g., an ester,anhydride, or an anhydride linkage.

Peptides are two or more amino acids joined by a peptide bond. Peptidescan vary in length from dipeptides with two amino acids to polypeptideswith several hundred amino acids. See Chambers Biological Dictionary,editor Peter M. B. Walker, Cambridge, England: Chambers Cambridge, 1989,page 215. Special mention is made of di-peptides, tri-peptides,tetra-peptides, and penta-peptides.

The carriers described herein may be derived from amino acids and can bereadily prepared from amino acids by methods within the skill of thosein the art based upon the present disclosure and the methods describedin WO96/30036, WO97/36480, U.S. Pat. Nos. 5,643,957 and 5,650,386. Forexample, the compounds may be prepared by reacting the single aminoacid, mixtures of two or more amino acids, or amino acid esters with theappropriate acylating or amine-modifying agent, which reacts with a freeamino moiety present in the amino acid to form amides. Protecting groupsmay be used to avoid unwanted side reactions as would be known to thoseskilled in the art. With regard to protecting groups, reference is madeto T. W. Greene, Protecting Groups in Organic Synthesis, Wiley, New York(1981), the disclosure of which is hereby incorporated herein byreference.

Suitable, but not limiting, examples of acylating agents useful inpreparing an acylated amino acids include the acid chloride acylatingagents having the formula

wherein R¹ is an appropriate group for the modified amino acid beingprepared, such as, but not limited to, alkyl, alkenyl, cycloalkyl, oraromatic, and particularly methyl, ethyl, cyclohexyl, cyclophenyl,phenyl, or benzyl, and X is a leaving group. Typical leaving groupsinclude, but are not limited to, halogens such as chlorine, bromine andiodine.

Examples of the acylating agents include, but are not limited to, acylhalides including, but not limited to, acetyl chloride, propyl chloride,cyclohexanoyl chloride, cyclopen-tanoyl chloride, and cycloheptanoylchloride, benzoyl chloride, hippuryl chloride and the like; andanhydrides, such as acetic anhydride, propyl anhydride, cyclohexanoicanhydride, benzoic anhydride, hippuric anhydride and the like. Preferredacylating agents include benzoyl chloride, hippuryl chloride, acetylchloride, cyclohexanoyl chloride, cyclopentanoyl chloride, andcycloheptanoyl chloride.

The amine groups can also be modified by the reaction of a carboxylicacid with coupling agents such as the carbodiimide derivatives of aminoacids, particularly hydrophilic amino acids such as phenylalanine,tryptophan, and tyrosine. Further examples includedicyclohexylcarbodiimide and the like.

If the amino acid is multifunctional, i.e., has more than one —OH, —NH,or —SH group, then it may optionally be acylated at one or morefunctional groups to form, for example, an ester, amide, or thioesterlinkage.

For example, in the preparation of many acylated amino acids, the aminoacids are dissolved in an aqueous alkaline solution of a metalhydroxide, e.g., sodium or potassium hydroxide and the acylating agentadded. The reaction time can range from about 1 hour to about 4 hours,preferably about 2 to about 2.5 hours. The temperature of the mixture ismaintained at a temperature generally ranging between about 5° C. andabout 70° C., preferably between about 10° C. and about 50° C. Theamount of alkali employed per equivalent of NH, groups in the aminoacids generally ranges between about 1.25 moles and about 3 moles, andis preferably between about 1.5 moles and about 2.25 moles perequivalent of NH₂. The pH of the reaction solution generally rangesbetween about pH 8 and about pH 13, and is preferably between about pH10 and about pH 12. The amount of amino modifying agent employed inrelation to the quantity of amino acids is based on the moles of totalfree NH, in the amino acids. In general, the amino modifying agent isemployed in an amount ranging between about 0.5 and about 2.5 moleequivalents, preferably between about 0.75 and about 1.25 equivalents,per molar equivalent of total NH, groups in the amino acids.

The modified amino acid formation reaction is typically quenched byadjusting the pH of the mixture with a suitable acid, e.g., concentratedhydrochloric acid, until the pH reaches between about 2 and about 3. Themixture separates on standing at room temperature to form a transparentupper layer and a white or off-white precipitate. The upper layer isdiscarded, and modified amino acids are collected by filtration ordecantation. The crude modified amino acids are then mixed with water.Insoluble materials are removed by filtration, and the filtrate is driedin vacuo. The yield of modified amino acids generally ranges betweenabout 30 and about 60%, and usually about 45%. The present inventionalso contemplates amino acids which have been modified by multipleacylation, e.g., diacylation, triacylation, etc.

If amino acid esters or amides are the starting materials, they aredissolved in a suitable organic solvent such as dimethylformamide orpyridine, and are reacted with the amino modifying agent at atemperature ranging between about 5° C. and about 70° C., preferablyabout 250° C., for a period ranging between about 7 and about 24 hours.The amount of amino modifying agents used relative to the amino acidesters are the same as described above for amino acids.

Thereafter, the reaction solvent is removed under negative pressure, andoptionally, the ester or amide functionality can be removed byhydrolyzing the modified amino acid ester with a suitable alkalinesolution, e.g., 1N sodium hydroxide, at a temperature ranging betweenabout 50° C., and about 80° C., preferably about 70° C., for a period oftime sufficient to hydrolyze off the ester group and form the modifiedamino acid having a free carboxyl group. The hydrolysis mixture is thencooled to room temperature and acidified, e.g., with an aqueous 25%hydrochloric acid solution, to a pH ranging between about 2 and about2.5. The modified amino acid precipitates out of solution and isrecovered by conventional means such as filtration or decantation.

The modified amino acids may be purified by acid precipitation,recrystallization, or fractionation on one or more solid chromatographicsupports, alone or linked in tandem. Suitable recrystallization solventsystems include, but are not limited to, acetonitrile, methanol, andtetrahydrofuran. Fractionation may be performed on a suitablechromatographic support such as silica gel or alumina, using solventmixtures such as methanol/n-propanol mixtures or aceticacid/butanol/water mixtures as the mobile phase; reverse phase columnsupports using trifluoroacetic acid/acetonitrile mixtures as the mobilephase, and ion exchange chromatography using water as the mobile phase.When anion exchange chromatography is performed, preferably a 0-500 mMsodium chloride gradient is employed. The modified amino acids may alsobe purified by extraction with a lower alcohol such as methanol,butanol, or isopropanol to remove impurities such as inorganic salts.

The modified amino acids generally are soluble in alkaline aqueoussolution (pH≧9.0); partially soluble in ethanol, n-butanol and 1:1 (v/v)toluene/ethanol solution; and insoluble in neutral water. The alkalimetal salts, e.g., the sodium salts of the modified amino acids, aregenerally soluble in water at about a pH of 6-8.

In poly amino acids or peptides, one or more of the amino acids may bemodified acylated and/or sulfonated. Poly amino acids and peptides mayinclude one or more acylated amino acid(s). Although linear modifiedpoly amino acids and peptides will generally include only one acylatedamino acid, other poly amino acid and peptide configurations can includemore than one acylated amino acid. Poly amino acids and peptides can bepolymerized with the acylated amino acid(s) or can be acylated afterpolymerization.

Delivery Systems

The compositions of the present invention may include one or more activeagents.

In one embodiment, the compounds above or salts of these compounds orpoly amino acids or peptides that include at least one of thesecompounds or salts may be used directly as a delivery carrier by simplymixing one or more compounds or salts, poly amino acids or peptides withthe active agent prior to administration.

The administration mixtures are prepared by mixing an aqueous solutionof the carrier with an aqueous solution of the active ingredient, justprior to administration. Alternatively, the carrier and the biologicallyor chemically active ingredient can be admixed during the manufacturingprocess. The solutions may optionally contain additives such asphosphate buffer salts, citric acid, acetic acid, gelatin, and gumacacia.

Stabilizing additives may be incorporated into the carrier solution.With some drugs, the presence of such additives promotes the stabilityand dispersibility of the agent in solution. The stabilizing additivesmay be employed at a concentration ranging between about 0.1 and 5%(w/v), preferably about 0.5% (w/v). Suitable, but non-limiting examplesof stabilizing additives include gum acacia, gelatin, methyl cellulose,polyethylene glycol, carboxylic acids and salts thereof, and polylysine.The preferred stabilizing additives are gum acacia, gelatin, and methylcellulose.

The amount of active agent is an amount effective to accomplish thepurpose of the particular active agent for the target indication. Theamount of active agent in the compositions typically is apharmacologically, biologically, therapeutically, or chemicallyeffective amount. However, the amount can be less than apharmacologically, biologically, therapeutically, or chemicallyeffective amount when the composition is used in a dosage unit form,such as a capsule, a tablet, a powder, or a liquid, because the dosageunit form may contain a plurality of carrier/biologically or chemicallyactive agent compositions of may contain a divided pharmacologically,biologically, therapeutically, or chemically effective amount. The totaleffective amount can then be administered in cumulative unitscontaining, in total, pharmacologically, biologically, therapeuticallyor chemically active amounts of biologically or pharmacologically activeagent.

The total amount of active agent, and particularly biologically orchemically active agent, to be used can be determined by those skilledin the art. However, it has surprisingly been found that with somebiologically or chemically active agents, the use of the presentlydisclosed carriers provides extremely efficient delivery, particularlyin oral, intranasal, sublingual, intraduodenal, subcutaneous, buccal,intracolonic, rectal, vaginal, mucosal, pulmonary, transdermal,intradermal, parenteral, intravenous, intramuscular, and ocular systems,as well as traversing the blood-brain barrier. Therefore, lower amountsof biologically or chemically active agents than those used in priordosage unit forms or delivery systems can be administered to thesubject, while still achieving the same blood levels and therapeuticeffects.

The amount of carrier in the present compositions is a deliveryeffective amount and can be determined for any particular carrier orbiologically or chemically active agent by methods known to thoseskilled in the art. The effective amounts of the active agent andcarrier in the composition may vary over a considerable range and isdependent upon the age, weight, sex, sensitivity, medical history, andthe like of the individual. Clearly, it is necessary to take intoaccount the nature of the active agent and carrier, the specificactivity of the agent (units of bioactivity/mass), and its rate ofabsorption in the gastrointestinal tract, all of which contribute to adetermination of the therapeutically effective dose.

Following administration, the active agent present in the composition ordosage unit form is rapidly taken up into the circulation. Thebioavailability of the ingested agent is readily assessed by measuring aknown pharmacological activity in blood, e.g. an increase in bloodclotting time caused by heparin, or a decrease in circulating calciumlevels caused by calcitonin.

Dosage unit forms can also include any of excipients, diluents,disintegrants, lubricants, plasticizers, colorants, flavorants,taste-masking agents, sugars, sweeteners, salts, and dosing vehicles,including, but not limited to water, 1,2-propane diol, ethanol, oliveoil, or any combination thereof.

The delivery compositions of the present invention may also include oneor more enzyme inhibitors. Such enzyme inhibitors include, but are notlimited to, compounds such as actinonin or epiactinonin and derivativesthereof. Derivatives of these compounds are disclosed in U.S. Pat. No.5,206,384, the disclosure of which is hereby incorporated herein byreference. Other enzyme inhibitors include, but are not limited to,aprotinin (Trasylol) and Bowman-Birk inhibitor.

The system is particularly advantageous for delivering chemically orbiologically active agents which would otherwise be destroyed orrendered less effective by conditions encountered before the activeagent reaches its target zone (i.e. the area in which the active agentof the delivery composition is to be released) and within the body ofthe animal to which they are administered. Particularly, the presentinvention is useful in pulmonary administration, such as by an inhaler,of active agents, especially those which are not ordinarily deliverableby that route or those for which improved delivery is desired. Improveddelivery may be faster delivery or greater delivery in a given timeperiod.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following examples illustrate the invention without limitation. Allparts are given by weight unless otherwise indicated.

EXAMPLE 1 Preparation of 2-(4-(N-salicyloyl)aminophenyl)propionic Acid

A slurry of 58.6 g (0.355 mol) of 2-(4-aminophenyl)propionicacid and 500ml of methylene chloride was treated with 90.11 ml (77.13 g, 0-710 mol)of trimethylsilyl chloride and was heated to reflux for 120 min. Thereaction mixture was cooled to 0° C. and treated with 184.44 ml (107.77g, 1.065 mol) of triethylamine. After stirring for 5 minutes, thismixture was treated with a solution of 70.45 g (0.355 mol) ofO-acetylsalicyloyl chloride and 150 ml of methylene chloride. Thereaction mixture was warmed to 25° C. and stirred for 64 hr. Thevolatiles were removed in vacuo. The residue was stirred in 2N aqueoussodium hydroxide for one hour and acidified with 2 M aqueous sulfuricacid. The solid was recrystallized twice from ethanol/water to give atan solid. Isolation by filtration afforded 53.05 g of (52% yield) of2-(4-(N-salicyloyl)aminophenyl)propionic acid.

Properties

Solubility: 200 mg/ml: 200 mg+350 μl 2N NaOH+650 μl H₂O-pH-7.67. Anal.calculated for —C,67.36; H, 5.3; N, 4.91. Found —C, 67.05; H, 5.25; N,4.72.

EXAMPLE 2 Preparation of Sodium2-(4-(N-salicyloyl)aminophenyl)progionate

A solution of 53.05 g (0.186 mol) of2-(4-(N-salicyloyl)aminophenyl)propionic acid and 300 ml of ethanol wastreated with 7.59 g (0.190 mol) of NaOH dissolved in 22 ml of water. Thereaction mixture was stirred for 30 min at 25C. and for 30 min at 0° C.The resulting pale yellow solid was isolated by filtration to give 52.61g of sodium 2-(4-(N-salicyloyl)aminophenyl)propionate.

Properties

Solubility: 200 mg/ml clear solution, pH=6.85. Anal. calculated for —C,60.45; H, 5.45; N, 3.92; Na, 6.43. Found —C, 60.84; H, 5.87; N, 3.85;Na, 6.43. Melting point 236-238° C.

EXAMPLE 3

A pulmonary delivery dosing composition of 0.1 mg/kg of porcine insulinand 7.5 mg/kg of the compound of Example 2 in water was prepared. AO.3ml/kg dose of the pulmonary dosing composition at pH 7.3-7.6 wasadministered to five normal, non-fasting rats by the followingprocedure. 1½ Popper and Sons gavage needles were inserted about a fewcentimeters down the animals' throats. The tip of the needed wasmanipulated towards the animals' ventral sides where the needle couldfall into a pocket and then with further manipulation the trachea. Oncethe needle was in the trachea, the dosing solution was delivered throughthe needle.

Periodic blood samples were drawn via the tail artery, and blood glucoselevels were measured using Ektachem DT slides (Johnson & JohnsonClinical Diagnostics, Inc., Rochester N.Y.).

Results are illustrated in FIG. 1.

Comparative Example 3A

The procedure of Example 3 was followed, substituting a dosingcomposition of 0.1 mg/kg of porcine insulin and water for the dosingcomposition.

Results are illustrated in FIG. 1.

Comparative Example 3B

The procedure of Example 3 was followed, substituting a dosing solutionof 7.5 mg/kg of the compound prepared in Example 1 in water for thedosing solution.

Results are illustrated in FIG. 1.

EXAMPLE 4

The procedure of Example 3 was followed, substituting a dosingcomposition of 0.5 mg/kg of porcine insulin and 7.5 mg/kg of thecompound prepared in Example 2, in water for the dosing composition atpH 6.6-6.9.

Results are illustrated in FIG. 2.

Comparative Example 4A

The procedure of Example 4 was followed, substituting a dosingcomposition of 0.5 mg/kg of porcine insulin in water for the dosingcomposition.

Results are illustrated in FIG. 2.

Comparative Example 4B

The procedure of Example 4 was followed, substituting a dosingcomposition of 7.5 mg/kg of the compound prepared in Example 2 in waterfor the dosing solution.

Results are illustrated in FIG. 2.

EXAMPLE 5

A dosing composition of 300 mg/kg of the compound prepared in Examples 2and 3 mg/kg of human insulin in water at pH 7.02 was prepared. Streptcatherized rats were dosed by oral gavage with 1 ml/kg of the dosingcomposition. Blood glucose levels (mg/dL) were measured.

Results are illustrated in Table 1 below.

TABLE 1 Time (hrs): 0 0.5 1 2 3 4 Assay: 0 −44.9 −42.59 −37.58 36.62−29.88 SD: 0 13.88 28.33 30.87 26.27 18.61

EXAMPLE 6

A dosing composition of 300 mg/kg of the compound prepared in Example 1and 100 Mg/kg of the N-terminal fragment of parathyroid hormone (aminoacids 1-34) in water at pH 8.13 was prepared. Normal rats were dosed byoral gavage with 1 ml/kg of the dosing composition. Serum levels ofparathyroid hormone (pg/mL) were measured.

Results are illustrated in in Table 2 below.

TABLE 2 Time(mins): 0 15 30 45 60 90 Assay: 0 1522.53 886.5 786.46 711.6482.46 SD: 0 538.5 272.21 239.78 181.65 329.78

EXAMPLE 7

A dosing composition of 100 mg/kg of the compound prepared in Example 1and 200 ug/kg of the N-terminal fragment of parathyroid hormone (aminoacids 1-34) in water at pH 7.65 was prepared. Normal rats were dosed byoral gavage with 1 mg/kg of the dosing composition. Serum levels ofparathyroid hormone (pg/mL) were measured.

Results are illustrated in Table 3 below.

TABLE 3 Time(mins): 0 15 30 45 60 90 Assay: 0 929.6 767.27 986.79 411.03356.8 SD: 0 693.18 525.18 1211.9 136.15 61.13

EXAMPLE 8

The method of Example 7 was followed but the pH of the dosingcomposition was 7.55.

Results are illustrated in Table 4 below.

TABLE 4 Time(mins): 0 15 30 45 60 90 Assay: 0 976.79 711.42 610.78520.51 516.19 SD: 0 388.07 244.89 218.16 78.31 37.6

EXAMPLE 9 Pulmonary Insulin Delivery

The carrier compound 2-(4-(N-salicyloyl)aminophenyl)propionic acid wastested as follows. Each rat was weighed and identified using anindelible marker and anesthetized by intramuscular injection ofthorazine (3 mg/kg) and ketamine (44 mg/kg).

An endotracheal tube (spray instillator from Penn Century ofPhiladelphia, Pa.) with a syringe (Hamilton syringe) attached to theendotracheal tube was inserted using a fiber optic laryngoscope. Thesyringe (Hamilton syringe) of the instillator was used to instillinsulin (0.03 mg/kg) and a carrier compound (16 mg/kg) into the lowerportions of the airway. The endotracheal tube was removed followingadministration, and the breathing rate was monitored throughout theremainder of the study.

Blood samples were withdrawn at 0, 5, 15, 30, 60, and 120 minutes viathe tail artery and assayed with a DSL Insulin Kit #10-1600 followingthe procedure outlined in the kit. The C_(max) value with the carrierwas 44.6±10.0. The C_(max) value without the carrier was 19.4±4.3.

EXAMPLE 10 Recombinant Human Growth Hormone (rhGH) Oral Delivery

Oral gavage (PO) dosing solutions containing 200 mg/kg carrier compound2-(4-(N-salicyloyl)aminophenyl)propionic acid and 3 mg/kg rhGH in 100%phosphate buffer were prepared by mixing. The pH of the solutions rangedfrom about 7.67 to about 8.09.

Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours andadministered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15minutes prior to dosing. The rats were administered 1 ml/kg of thedosing solution by PO. Blood samples were collected parenterally fromthe tail artery for determination of serum rHGH concentrations. SerumrHGH concentrations were quantified by an rHGH immunoassay test kit (Kit#K1F4015 from Genzyme Corporation Inc., Cambridge, Mass.). The mean peakserum concentration of rhGH was 93±74 ng/ml.

EXAMPLE 11 Erythropoietin (EPO) Oral Delivery

Oral gavage (PO) dosing solutions containing 200 mg/kg carrier compound2-(4-(N-salicyloyl)aminophenyl)propionic acid and 1 mg/kg humanerythropoietin quantikine IUD in 100% water were prepared by mixing. ThepH of the solutions ranged from about 6.9 to about 7.9.

Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours andadministered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15minutes prior to dosing. The rats were administered 1 ml/kg of thedosing solution by PO. Blood samples were collected parenterally fromthe tail artery for determination of serum EPO concentrations. Serum EPOconcentrations were quantified by an Elisa assay test kit (Kit #DEPOOfrom R&D Systems, Minneapolis, Minn.). The mean peak serum concentrationof EPO was 163±136 ng/ml.

The procedure was repeated without the carrier compound. The mean peakserum concentration of EPO was 0 ng/ml.

The above mentioned patents, applications, test methods, andpublications are hereby incorporated by reference in their entirety.

Many variations of the present invention will suggest themselves tothose skilled in the art in light of the above detailed description. Allsuch obvious variations are within the full intended scope of theappended claim.

What is claimed is:
 1. A salt of the compound having the formula

wherein the salt is not a sodium salt.
 2. A composition comprising: (A)at least one active agent; and (B) a carrier comprising a compoundhaving the formula

 or salt thereof.
 3. A composition as defined in claim 2, wherein saidactive agent is selected from the group consisting of a biologicallyactive agent, a chemically active agent, or a combination thereof.
 4. Acomposition as defined in claim 3, wherein said biologically activeagent comprises at least one peptide, mucopolysaccharide, carbohydrate,or lipid.
 5. A composition as defined in claim 3, wherein saidbiologically active agent is selected from the group consisting ofgrowth hormones, human growth hormones (hGH), recombinant human growthhormones (rhGH), bovine growth hormone, porcine growth hormones, growthhormone-releasing hormones, interferons, α-interferon, β-interferon,γ-interferon, interleukin-1, interleukin-II; insulin, insulin-likegrowth factor(IGF), IGF-1, heparin, unfractionated heparin, heparinoids,dermatans, chondroitins, low molecular weight heparin, very lowmolecular weight heparin, ultra low molecular weight heparin,calcitonin, salmon calcitonin, eel calcitonin, human calcitonin,erythropoietin (EPO), atrial naturetic factor, antigens, monoclonalantibodies, somatostatin, protease inhibitors, adrenocorticotropin,gonadotropin releasing hormone, oxytocin,leutinizing-hormone-releasing-hormone, follicle stimulating hormone,glucocerebrosidase, thrombopoietin, filgrastim, prostaglandins,cyclosporin, vasopressin, sodium chromoglycate, disodium chromoglycate,vancomycin, desferrioxamine (DFO), parathyroid hormone (PTH), fragmentsof PTH, antimicrobials, anti-fungal agents, analogs, fragments, mimeticsand polyethylene glycol (PEG)-modified derivatives of these compounds;and any combination thereof.
 6. A composition as defined in claim 3,wherein said biologically active agent is selected from the groupconsisting of human growth hormones (hGH), bovine growth hormone, growthhormone-releasing hormones, interferons, interleukin-1, interleukin-II,insulin, heparin, low molecular weight heparin, very low molecularweight heparin, calcitonin, erythropoietin (EPO), atrial natureticfactor, antigens, monoclonal antibodies, somatostatin,adrenocorticotropin, gonadotropin releasing hormone, oxytocin,vasopressin, sodium chromoglycate, disodium chromoglycate, vancomycin,desferrioxamine (DFO), parathyroid hormone (PTH), antimicrobials,anti-fungal agents, and any combination thereof.
 7. A composition asdefined in claim 3, wherein said biologically active agent comprises aninterferon, interleukin-II, insulin, heparin, low molecular weightheparin, very low molecular weight heparin, calcitonin, parathyroidhormone (PTH), erythropoietin (EPO), human growth hormone (hGH),oxytocin, vasopressin, vancomycin, desferrioxamine (DFO), parathyroidhormone, and combinations thereof.
 8. A composition as defined in claim3, wherein said biologically active agent comprises unfractionatedheparin, ultra low molecular weight heparin, and combinations thereof.9. A composition as defined in claim 3, wherein said biologically activeagent comprises insulin.
 10. A composition as defined in claim 3,wherein said biologically active agent comprises parathyroid hormone.11. A composition as defined in claim 3, wherein said biologicallyactive agent comprises human growth hormone.
 12. A composition asdefined in claim 3, wherein said biologically active agent compriseserythropoietin.
 13. A composition as defined in claim 2, wherein saidcarrier comprises a poly(amino acid) or a polypeptide.
 14. A dosage unitform comprising: (A) a composition as defined in claim 2; and (B) (a) anexcipient (b) a diluent, (c) a disintegrant, (d) a lubricant, (e) aplasticizer, (f) a colorant, (g) a dosing vehicle, or (h) anycombination thereof.
 15. A dosage unit form as defined in claim 13,wherein said active agent is selected from the group consisting of abiologically active agent, a chemically active agent, or a combinationthereof.
 16. A dosage unit form as defined in claim 14, wherein saidbiologically active agent comprises at least one peptide,mucopolysaccharide, carbohydrate, or lipid.
 17. A dosage unit form asdefined in claim 14, wherein said biologically active agent is selectedfrom the group consisting of growth hormones, human growth hormones(hGH), recombinant human growth hormones (rhGH), bovine growth hormone,porcine growth hormones, growth hormone-releasing hormones, interferons,α-interferon, β-interferon, γ-interferon, interleukin-1, interleukin-II,insulin, insulin-like growth factor(IGF), IGF-1, heparin, unfractionatedheparin, heparinoids, dermatans, chondroitins, low molecular weightheparin, very low molecular weight heparin, ultra low molecular weightheparin, calcitonin, salmon calcitonin, eel calcitonin, humancalcitonin, erythropoietin (EPO), atrial naturetic factor, antigens,monoclonal antibodies, somatostatin, protease inhibitors,adrenocorticotropin, gonadotropin releasing hormone, oxytocin,leutinizing-hormone-releasing-hormone, follicle stimulating hormone,glucocerebrosidase, thrombopoietin, filgrastim, prostaglandins,cyclosporin, vasopressin, sodium chromoglycate, disodium chromoglycate,vancomycin, desferrioxamine (DFO), parathyroid hormone (PTH), fragmentsof PTH, antimicrobials, anti-fungal agents, analogs, fragments, mimeticsand polyethylene glycol (PEG)-modified derivatives of these compounds,and any combination thereof.
 18. A dosage unit form as defined in claim14, wherein said biologically active agent is selected from the groupconsisting of human growth hormones (hGH), bovine growth hormone, growthhormone-releasing hormones, interferons, interleukin-1, interleukin-II,insulin, heparin, low molecular weight heparin, very low molecularweight heparin, calcitonin, erythropoietin (EPO), atrial natureticfactor, antigens, monoclonal antibodies, somatostatin,adrenocorticotropin, gonadotropin releasing hormone, oxytocin,vasopressin, sodium chromoglycate, disodium chromoglycate, vancomycin,desferrioxamine (DFO), parathyroid hormone (PTH), antimicrobials,anti-fungal agents, and any combination thereof.
 19. A dosage unit formas defined in claim 14, wherein said biologically active agent comprisesan interferon, interleukin-II, insulin, heparin, low molecular weightheparin, very low molecular weight heparin, calcitonin, parathyroidhormone (PTH), erythropoietin (EPO), human growth hormone (hGH),oxytocin, vasopressin, vancomycin, desferrioxamine (DFO), parathyroidhormone, and combinations thereof.
 20. A dosage unit form as defined inclaim 14, wherein said biologically active agent comprisesunfractionated heparin, ultra low molecular weight heparin, andcombinations thereof.
 21. A dosage unit form as defined in claim 14,wherein said biologically active agent comprises insulin.
 22. A dosageunit form as defined in claim 14, wherein said biologically active agentcomprises parathyroid hormone.
 23. A dosage unit form as defined inclaim 14, wherein said biologically active agent comprises human growthhormone.
 24. A dosage unit form as defined in claim 14, wherein saidbiologically active agent comprises erythropoietin.
 25. A dosage unitform as defined in claim 13, comprising a tablet, a powder, a capsule,or a liquid.
 26. A method for administering a biologically-active agentto an animal in need of said agent, said method comprising administeringorally to said animal a composition as defined in claim
 3. 27. A methodfor preparing a composition comprising mixing: (A) at least one activeagent; (B) the compound of claim 1; and (C) optionally, a dosingvehicle.